CA2036107A1 - Contrast media synthesized from polyaldehydes - Google Patents

Abstract

Abstract Microparticles comprising biodegradable polymers, characterized in that they are synthesized from polymerizable aldehydes, which optionally contain additives and/or crosslinkers capable of copolymerization, optionally surfactants or surfactant mixtures, gases and/or highly volatile liquids in free or bound form, coupling agents, optionally biomolecules or macromolecules bound by these coupling agents as well as optionally suitable as ultrasonic contrast media.

Description

~Q ~r ~
Th~ inv~n-kion r~l~te~ to n~ mi~rop~icles, ph~rmaceutlcal ag~n~ aontaining them, ~he~r u~e, e.~., in ultrasonic diagnogis as w~ll a prooesse~ for the p~odu~tion o~ the~ microparticles and pha~maceutic~l ayents.
It iG known that by pe~ipheral injeotion of solutions, which ~ontain fine ~as .bubbles~ oardiac echo çontra~ts can ~e achieved (Roelandt JO~ Ultra~ound Med~ Biol. 8: ~71-4g~,1982). T~e6e air bubble~ are ob~ained in phy iologically c~mpatible ~olutions, e.g., by ~h~king, other a~itation or ~y addi~ion o~ car~on divxide. But kh~y ~re n~ uni~orm in ~egard t~ number ~nd siæe~
and ~an b~ reproduc~d only un~atis~c~orily. Al~o ~h~y generally ar~ no~ sta~ z~d so ~hat their life is ~hort. Their av~rage diameter~ in mo~t case~ above th~ ~ize of erythroc~te~ so that no lung capill~ry pasOEag~ with ~ubse~uent ~ontrast1n~ of o~gans ~u~h ~ le~t hear~, liver, kidney or ~pleen i5 po~ le.
Furth~r, they are not ~uitable for ~ antl~iaatlon3, ~ince ~he ultr~ound echo produced by th~m i~ composed of ~ev~r~l in~eparable pro~e~sss such a~ ~ub~le ~ormation, coale~enc~ and : di~olution, ~hu~, ~.g., lt i not possible wi~h the~e ultras~niG con~ra~ ~edia to o~tain di~p~ays on tran~it time~ by ~th~ ~a~urefflen~ ~ th~ contra~t cour~e in the myocardium.
`~ The sta~illzation o~ ~he g~ bubblea by sugar is des~ribe~
ln EP O 131 540. Thu~ ~he reproduoibility and homoge.neity of ~he COD~ra8t ef~e~t i~ îndeed im~roved but th~se bubble~ do not r~ lung pa~1bag~.

It i~3 descrlbed in ~P o 122 6~4 and o 123 235 ~h~t the gaa bubble s~ iliæing e~fect of ~uga~;, 3ugar alcohol~: and ~alts i3 imprc)~ed ~y adflition of 3urfactant~. A l~ln~ capillary pa~age and the po~sibility of ~epres~ntation of the a~terial ves~el branch and di~erent org~n~ ~uch as l iver a~ld ~plQen exists with thlæ ultra~onic c:~n~ra~-t. media. But here -~he contra~ e~ect is limited ~o ~he vQssel lumet~, ~ince the bubble~ are not absorbed }~y ~h~ ti~ue aell~.
Nc~ne of th~ ultrasonic: con~rast m~aia known so ~ar rem~ins unch~ngRd ~or a ~rolon~ed period ~ n ~he l~ody . Therefore an organ rRpresentation wilth ~uf~i~ierlt signal inten~lty ~y ~el~ctive COhce3ntration after i.v. in~ecltion or ~uanti~iq~tion~ ara not pOS~ Q.
An ~nc~psulation of g~e6, such 21s, ~or exampl~, air as ul~ra:3onic contr~st medi~ de~arihed in DE-OS 3~ 03 ~72. Th~
wall materi~l u3ed here con~ifi~qs oi~ biodegrad~ble ~;ynthetic ma~erial, E;uch ~s e~pe ;:ially cyanoacryla~e and polylactide~
BU~ the~e mi~roparti.cle~ -- e~peal~lly on larg~r ~c~le as w 11 as in re~ar~ to their working up -- are difficUlt to produ~. Thus ~pecially the bro~d BiZ~ t~i~ution o~ these particl ~ 1~ a drawbaak.

This lnven~ion providQ~ ul~rasoni~ ontra~t medla, which do not exhihit ~hes~ draw~ack~ and i~ achieved by th~ p~oYisior of ~he micropart~cle~ a cord~ng ~o ~he invention~
The~e mi~ropar~i~le; con i~t of biod~gradable polymers, chaxac:teriz~d in ~h~t th~y are ~ynthe~ized from polymeriza~le 2 ~

ald~hyde~ which optionally aontain additive~ and/or crosslinkers aap~ble of ~op~lymerization, op~lonally surfac-tant~ or ~urfaatant mixt~re~, g~e~ and~or highly volatile li~l~d~ in free or bound f~rm, coupling ~gent~, optionally biomoleculQY or macromolecule~
bound by theg~ coupling agents as well as optionally diagno~tic~lly or therapeutically e~fectivR co~ponents.
In general, thq polymerizable aldehyde~ can be selec~ed from a wid~ valiety o~ R-CHo g~oups, wher~in R 1~ non-crit~oal and can ~e ~elected ~roml e.g., cl 10 hydrocarbons whi~h c:an b~ ~aturated ; or un~turated, acy~lic or cyclic, aliphatic or aroma~ic, op~i~n~lly sub~tituted by h~lo, other eu~h hydrocar~on group~, : CN, CHO, etc . Pr~f ~rxed are alkane aldehydsa, acrol~n derivative~ ~nd dlaldehyd~s.
sult~ble polymeri~ed aldehyde~ use~ul in the syn~lle~i~ o~
~the micropartioles can ~e sele~t~d ~rom the following exe~plary preferred polym~riz~ble aldehyde monomar~:
I. alpha,be~a-unsatura~ed aldehyde~, sU~h a~, ~or ~xampl~

~`. acroleln : cro~on~ldehyde pr~pynoicald~hyde alpha~~ub~ti~u~ea aGrol~in derivativ~s, ~u~h a~, f~r example alpha-m~hyl aarolein lpha~hloroac~olein alpha-ph~yl aaroIein alpha-ethyl acrolein alph~-i60propyl a~role~n alpha~n-but~l 4~role~n ~lph~-n-pr~pyl acroleiD

2~3~

III. ~laldehyd~ ch a;, for example glu~ald~-hyde, succinaldehyde or their deriva~ a~ or ~heir mlx~ur~s wi~h additive~ ~pable of copolymexization ~co~nonom~r~
~uah as, ~or ~xample alpha-~qubstitu~ed a~roleins b~ta-s~s~ltute~ acroleins ethyl ayano~cryla~q6 methyl cyanoacry 1 ate~
bu~yl c:yanoacryl~te~3 h~xyl c:yanoa~ryl a t~
m~thyl me~hacrylate~
vinyl al~oh~l~
acrylic acidcL
m~thac:rylic 2~oid~
acryl i~ acid chloride~
m~th~qrylic ~c~id chloride~
a ary~onitri l e methacrylon~ triles acJ~ylamides sulo~tituted a~rylamide~
hydr~xy me~byl methacrylate~
mesityl oxid~
dime~hyl~minoethylmetha~ tes 2-vinylpyridine~
N-~rinyl -2 -pyrrol id ~none In thi~ e, ac~rolein and glutaraldehyda are pr~errE~d.
Comonomer~ o~ any of the ~ore~oing aldehydes with eac:h other ~nd with the oth~3r ~omonomer men~ioned ~bove ar~ also included.
In g~n~r~l, the amount o~ a~dehyd~ monom~r in the ~opolymer~ i~
50-1~0 m~l~ %, pref~r~bly 75-loO mol~ ne remaind~r b~ing the non-aldehyde. Where ~11 comc~nomer~ are ~ldehyd~s, ~he relative a~nount~ ~re not c:riti~al, ~ . g., ~ny c~ar3 l~e present ~n an amount ~ 0-100 mols %~

6 2~3~ ~7 All polymers (homopolyaldehyde~ or copolyald~hyde~) o~ thi~
invention typically h~ve mole~ular weig~ts o~ 1,000 d to 30,000 ~, prefar~bly 1,000 d to 12,000 d. 'rhe particle~ are hiodegr~da~le in hlood and pla~ma~
The ~ur~actan~ op-tio~ally involved in the ~ynthesis of the micropar~.lcle~ can be selected non-~r~tlcally from ionic (~atio~ ox anionlc) or nonionic surfaae-active 6ub3tance~
(~urfact~nts), su~h a~, e.g.
polye~hylene oxide polyoxyethylene polyoxypropylene~ ~uc:h a~
Pluronic~R) F fi8, Plu~nic~ F 108, Pluronic~R) F 1~7, polye~hylen~ glycol, Polox~mirl ~0~, Polaxamer 407 ~a~boxylic a~id ~alts, ~u~h ~s, ~or exampl~, sodium oleate p~lyoxyethylene f~t~y ~cid ~ers, su~h a~, f~r ex~mple:
p~lyoxy4thylene st~rat~
~odium di~yl 3ul fo~u~cinate polyglutar~ld~hydR sodium hydrogen~ul~i~e adduct polyacrolein sodium h~drog~n~ul~ite adduct polyvinyl ~ul~oni~ acid.
They can be Use~ alone or in the form ~f ~h~ir mixture~, the relative amount~ bein~ non-criti~al.
Pr~rred of these ~re~ p~lyglutaraldehyd~ ~odium ~ulf~te ad~uc~, polyacrolein ~odium ~y~ro~en~ulfite adduct, PlurQnia~
F68, Pluroniat~1 F 108 ~nd Pluronî~ F 1~7~
I~ the polymeriz~ble ~lde~yde~ use~ for the ~ynthe~i~ o~ t~e mlcrop~rti~ thems~lves have surfaatant prop~rtie~, th~n ~u~ tants may not ~e ~sed, bUt alsv can be ~ed. A~ an ex~ple of ~u~n ~ld~hyde~, glutaral~e~y~e Can ~ mentioned. ~ypiaally, the am~Un~ o~ ~ur~act~nt rel~ive ~o ~h~ amount of polyaldehyde i~ 0.0~-1 W~.~ and pre~ex~ed ~Ur~aae ~ension value~ for the ~ur~a~an~ ~ro 40-50 mN/m.
As ga~e~ o~ highly vola~ uid~ ~ontaln~d i~ the micro-~artiale~ ~ h ~r~e ~r bound form - in ~his c~s~, li~uids with ~oiling ~int b~low ~0~ ~re pre~erred ~ er~ are guit-abls phy~iolo~icall~ accepta~l~ ones ~a~ ~he minor amounts u~ed) ~u~3h a;:

2~3 am~nonia a~r lnert ga~e~ (heliu~, rlQon, argon, xenon, krypton~
sul~ur halid~s, ~ch as, ~or example: ~:ulfur hexafluoride nitrogen carbon oxid~s oxyg~
hy~rogen Hydrocarbon~ ~r thRir mix~Ure~, ~uch a~;, for ex~mpla:
m~th2~ne ethan~ -prop~nR
butane pentane neopen~ane isopent~ne cyclopen~ne ethylene propylerle ~c~ylsn~

3, 3 dimethyl-1-butyne ~, 3-p~n~adi~ne 2 -me!thyl -2 -butene 2-meth~l-1, 3-butadisrle ~-bukyne 2-~nethyl-1-b~ ne 3 -m~thyl - 1 -but~nq 2 ~ 3 ~

halo~n~t~d hydrocarb~ns or ~ixture~, such a~, Por exa~p~ ~:
methylQne P}IlC~rid~
1,l-dichloroRthylene i~opropyl ~hloridQ
dibr~modi~luorometh~ne bromom~thane e~hex~, ~uch a~, ~or exampl~: ~imethyl ~-ther, dieth~l ether ~ or ~luorinated e~h~r~, or ~ompoun~ su~h a~, Xor ~xample:
dim~thyl~mino~aeton~
propylene oxide N-ethyl~thylamln~
N-ethyldim~thyl~ine ~U~An ;~
Of the~e, air, ar~on, xenonr ~ulfur hexa~luoride, propane, butane and ~uran a~e pref~rred.
By "fr~" in ~his conkext i~ m~n~ that the ~a~ ar vol~til2 liguid ~s cont~ined in the microparticla by phy~i~al t~pping wh~r~a~ by ~ound" i8 meant a trapplng by weak ~hemical ~onding ~hort oS ~oYalent b~nding~ Typical ~ount~ relative ~o ~he amaunt of polymer~ are ~.01-2 wt.% ~or bQth ~he qa~ and volatile llquid~.
As coupl~ng agents utilizabl~ in the ~ynthe~is of micropar~l~le~ ~h~re are espe~ially ~uitabl~:
ompound3 ~or.taining amino groups, ~ch a~, ~or exampl~:
hydr~xyl amine 9 2~3~

bu t y :l arr~ 0 al lylamine ethanolamine trishydroxymethylaminometharle 3-alllino-1-plopallesulfonic aci~
5-aminovaleric acicl ~-aminooctanoic acid D-glucosamine hydrochloride aminogalactose aminosoIbitol aminomannitol diethylaminoethylamine anilines sulfonyl acid amide choline N-methylglucamine piperazine 1,6-hexanediamine urea --hydrazine glycine alanine lysine serine valine leucine -~3 ~L ~ 7 )tides proteins ,1 lbumin h u m;l n st~ r u m a 1 b um i n pol~ 1y~
~rt,~llltill polyglycolamin~s a~ninol)oly.llcoho Is de.~tran sulfates with amino groups N-aminopolyethylene glycol (I{O-PE(i-~'112) N,N'-diaminopolyethylene glycol (Nl{"-PE~-N~12) antibodies immunogloblllins II. Compounds containing acid groups, such as, e.g., carboxylic acids acetic acid propionic acid butyrlc acid valeric acid caproic acid caprylic acid capric acid lauric acid myristic acid palmitic acid stearic acid :
-~3 oleic a~id linoll~ aaidlinolenic acid cyc:lohexane c:~rboxylic acid phenylacetic acid benzoyla~eti~ id chlorobRnzo~ ~, acid ~romobRn~oic aci d n~tr~nzoio acid ortho-ph~hali~ a~id ~eta-ph halic a i~
p~ra-phthalic ~c:id ~alic:yl.ic aoid hydrox~enzoic ~cid aminob~rlzoi~;: acid ~e~hoxybenzoic acid OOH
PEG-NH~ O-CH,,CH2-~O-,NH-~H
CHz~ H
(PEG linX~r a~p~raginic acid~
PEG-lin~r-glu~aminia ~aid ~; PEG-îinkar-DTPA
- PEG~l ~ nker-ED~rA
III~ C!~mpc~und~ aont~ining hydroxy group6, ~uch a~, Pc~r Qxample:
al~ohol~
Jneth~nol 2 ~ 3 ~

ethano l propanol butallol pen-tano:l ~C',`i<lllO I
heptanol oc-tanol decanol dodecanol tetradecanol hexadecanol octadecanol isopropyl alcohol isobutyl alcohol isopentyl alcohol cyclopentc nol cyclohexanol crotyl alcohol benzyl alcohol ~phenyl alcohol diphenyl methanol triphenyl methanol cinnamyl alcohol ethylene glygol 1,3-propanediol glycerol 2~
pentaerythri-tol IV. Polymeri~able substances, such as alpha,beta-unsa-tnlated aldehy(les, s-lch as, e.g.:
acrolein cI~o~oll~llclellycl~
propiona:l.clebycle a].pha-.substituted acrolein derivatives, such as, e.g.:
alpha-methylacro].eirl alpha-chloroacrolein alpha-phenylacroleir : alpha-éthylacrolein : alpha-isopropylacrolein alpha-n-butylacrolein alpha-n-propylacrolein ; dia.ldehydes, such as, e.g.:
glutaraldehyde, suocinaldehyde or their derivatives or their mixtures wlth additives capable o~ copolymerization, such as, e.g,:
: :alpha-subs~tituted acroleins : ~ ~ beta-substituted acroleins ethyl cyanoacrylates methyl cyanoacrylates butyl acrylates hexyl cyanoacrylates methylmethacrylates 2 ~
vinyl alcohol~
a~rylic acids m~ha¢ryli~ ac~d~
acrylic acid chlorlde~
acxyl~nltril~.
~ethaaxyloni~riles a~ryl~mide~
~ub~titut~d ~cryl~mide~
hydroxym~thylmeth~ryla~e~
meaityl oxida dimethylaminoethylmathacrylate~ 2-~lnylpyridine ~-vinyl-2-pyrrolidinon~
: 0~ ~h~ th~ are preferred: hydro~yl~mine, tri~hydrox~me~hyla~inom~thane, 3-amino-l-propane ~ulfonic a~id, ~-glua~minohydroahloride, aminomannitol, urea, human serum albumin, hydrazine, prot~in~, polygly~lamine~, ~minopolyalcohol~
~u~h a~, ~.g., HO-PEG-NH2 or NH2~PEG-NH2 or compounds cont~lning a~id gr~up~ ~uah a~, e~g., PEG-link~r-a~pAraginic ~id, PEG-linker-gluta~inl~ acid, PE~}-link~r-DTPA and PE~-link~r~ q~, and thc moleaul~r weight o~ t]~ polye~hylenaglyc~l (Pi~ up to 1~0, ooO d, prefer~Iy unde~ ~10, ooo ~. Th~ ~mo~mt o~ aoupling ~gen~ pic~ly, base~ on th~ ~mount o~ poly~ldehyd~ 95 w~
The aoupling agent~ nam~d under X can be con~ens~d ~y ~nelr amlno group on th~ ~ormyl group~ loaated on the sur~a~ of micropart~cle~ ~ynthesizad ~r~m poly~e~ized al~ehyda~ and op~i4nally ~ur~tan~.

2 ~ 7 Also the monome~s ].istRd under IV, ~.~hich can be polymerized with other monome~ an be bound by the ~o~yl groups.
~ n ~h~ oth~r hand, ~he ~id~ and ~l~o~lols nam~d under 1~ ~nd III a~e typically coupled ~n ~he microparticles only ~fte~
pr~Viou~ conven~ional conveXsion of the aldehyde ~unction, e.g., as ~ cu~ed ~ow.
By th~ selea~lon o~ suita~le ~iomolecules or macromolecules, ~OUtl~ by these ~oupling ~g~n~S, ~uch a~, e.g., enzyme~, dex~rans, immunoglobulins, monoclonal an~ibodie~ (see belo~, micropa~tiGles aac~r~ing to the invention are ob~ained, which ex~i~it a urpri~ingly h~g~ sue and organ ~pecificity.
The ~icroparti~les accordin~ to the inven~lon optionally ~an contain dlagno~tically vr therapeutlcally eff~ctive components ~or diagnosl~ an~ therapy o~ ~umor~ ~uch as, ~or ex~mpl~
~ox~rubicin actinomyc~n m~gna~lt~
mitomycin C
triamcinolone, amon~ m~ny oth~rs, ~ypi~ally in amoU~ts up to ~0 ~t.9~ based on the ~mount o~ po.lyaldehyd~3, pre~i~e amounts b~ing routinRly dete~inable in a.~c:ordarlc~ wi~h typical do~ages ~or the ac:tive ~gf~nt And oth~r ronventl~nal considera~ions.
~ he ~nit;:ropar~is3le~i o~ ~he pre8en~ inv~n~ion are 8urprl ~ingly bet~er oompa~i~le esp~cially ~ rl vi~w of a~diova~cular ef ~ec~
(~locd pr~ure, hear~ rhyth~ia) and the p~rtic~ how no thrQmbogen ic po~ential .
Tne micropar~lcles aG~ording to the invention eXhi~it the de~cribe~ de~ired prop~rti~. T~ey can be produced imply and with ~i~h yi~l~. An up~c~ng o~ their production, a~ well as their purif~ation, i~ no probl~.
A~ cros~linker~ u~eful ~n ~hi~ in~ention axe includ~d the ~ompon~n~ listed belo~ a~ bi~unc:~ional linkers; typia~l amount~
are 0-5 wt . % . comonome~s ~nd aros~linker~ are typically $nc:1uded ~o f~nh2ln~e the ~tabili~y of th~ mi~roparti~les.
~h~ partlcl~s ~xhibit a n~rrow size dis~ri~ution (monodisper~); at the ~;~me ime, 1~ i~ possible t~ v~ry tha ~;ize 16 2~ P~

of the particles ove~ a wide range depen~lng on tha concentratl~n o~ the start~ng materials u~ed (~ee below). By contr~l of th~
produ~tion conditi~ns (~.g., pH) it ls pos~ible ~l~o to vary the molecular weight in wide ran~eq. Shape~ are no~ critical and ~ypically ar~ sphere~ ce.
A~other advan~ge ~onsi~ts in the faot that the rea¢tion for the ~ynt~e~i3 o~ the mi~roparti~ can be brought ~bout by many po~sibilltie~ known in prin~ipl~ for polymeriza~ion of aldehyde~, for example, an~onic polymeri~ation by change o~ pH, ~ationlc polymeriza~i~n, e.g., wi~h iron salt~, radl~l polym~rization wl~h W ligh~ and by ionizinq radla~ion.
The broad te~p~ratur~ range t-5 ~o +80~), possible for the production of the microparticles, allows a 8imple ~xperimental control Wi~h opti~al ylelds with very different ga~e~ or low-~o~ling li~Ulds.
The particle~ ~ntain ~ree aldeh~de ~roups, which ~an ~e ~ovalently lin~Qd to other ~le~u~es ~y ~hem~cal reaction8~ e.g.
a~ de6~cribed abc~e ~or coupling a~ents. Thi6 pos~ibili~y allows the proper~ies of the p;~rticle 3urfaae to bQ deliberately char~ged with~u~ a~eating th~ ~chogenic prvpertie3. The colloidal ~bility ~an b~ affected ~y ~he ~el~tlon o~ ~uitable molecules a~ coupling agent~. E3pec.ially ~n thi way ~he phenomenon of ~ggl~meration often occurring for ~olloidal sy~tem~ o~n be prev~nted. This ag~in i~ ~ gr~a~ a~vant~ge for th~ development o~ a ~table f4r~ulation.
~ e~ide affeating the ~tabili~y, po~ibilitie~ ~re of~ered of r.h~nging th~ ~urface o~ th~ par~iale8 ~ ~hat ~ drug targeting i~ po ~ . Thlg o~Ur3 hy coupling on o~ ~uitable biomolecule~
or m~aromole~ul~ (e~g4, m~noclonal anti~4d~), whi4h ~au~e a high ti~Ue ~nd ory~n ~pe~ificity (G. Gregoriadi~, G. Po~te, "Targeting ~r Drugs," Plenum Pre~ 8~, New ~ork) or by ~f~ectin~ the ~ur~ace proper~ies of th~ par~icl~ by absorp~on o~ m~l~sule~ ~e.g., ~urf~ct~nt~.
As ~ ~unctlon o~ the ~le~tion o~ th~se moleoule~ and o~ ~he 8i2~ 0~ the ~icrop~rti~le6, a parti~le ~on~n~r~ion ~an ~e 2 ~ 3 ~
achieve~ in/on tumors or, e.y., in the lung, liver, ~pleen and in ~he hone m~rrow. The concentration ln the bone marrow 1~
e6pe~1ally aahiev~d in that ~mall particle~ (le~3 than 100 nm) are coated, e.g., with Poloxamer 407. Xf the~e partlcles, e.g., ~re co~ted wlth P~loxamin 908, th~ REs i8 overcome ~y th~se partiale~, ~n~ th~y re~nain in th~ blood ci~ulatlon ~blood pool a~ent). Typical coati~lg thi~kn~s~es ar~ up to 2 nm.
~ y p~rti~ oupl~.d with anti~odie~, ~ concentrati~n of the parti~ in/on ~umors can b~ aohieYed.
An aatlve targeting can al~o b~ p~rEorm~ W~h mi~roparticles containin~ magnetite. T~e particle~ are concantra~ed in the d~ired places in the intrava~al ~ys~em by an externally applied ma~ne~ field. The pos~ibility to ~mre~tiga~e oir~ul~tory aondi~ions, e.g., ln the blood ve~el~, i8 thus ~s~hieved.
With th~ help of particle~ laden with magnetit~ it i5 a1~D
po~ible to produc:e lc~ lly hlgh temperature~3 by ~n ~x~ern~lly ir~dla~ed ~l~e~atlng magn~tic field. Thi~ an be u~d th~rap~utically, ~.g., ~r the d~tructiDn o~ tumor~ (hyperthermy thar~py)~ ("Hypa~he~ , Overgaard, ~., Th~ current and pot~nt~al rol~ of h~per~hexmi~ ln r~dio~erapy. Int. J. P.~dlatO
Oncol. Biolf Phy~ 1989, 1~:53$-54~ n ultrasoni~ ld ~n al~o be u~ed, be~id~s the uge of a maqnetia alternating ~leld.
As a result, at ~he ~ame ki~e ~trong l~aal ~emp~ra~ur~ increase~
~lea aGaur. (wo 89~051~0 - use o~ ~edia ~n~alnln~ bub~le~ in 3ho~kwav~ ~nd ul~ra~onic ther~py.) 18 2 ~ 3 ~ 3 7 Th~ product:i4rl of t~l~ micropa~t~ s ~ ordlng to the ~nv~n~on c:an tak~ plac~ in tha~ Wi~h s~irrlng ~n ~eous ti~n C:on~aining 0 ~o ~0%, pre:eer~l~ly 0.01 to 10% w~v of ~u~fa~tfln~(~) and o ~o 10% w~v o~ diagno~ all~ or th~r~peutically o~ecti~ compon~nks and yas~s or highly volatlle liquids~ at a temperature o~ -5~C to -~oVc~ preferably 0C to 4 0~C , a pH o~ 7 to 14, pre~3rably 9 ~o 13, within 1 minut~ to 24 hour~, pr~fe~a~ly 1 hou~- to 1o hour~, and op-tionally With t~e pa~age o~ ga~ ~herekhrough, i~ reac~ed wi~ copolymerizable ~ld~h~ds ~) up t-a a c~ncc3~tration r~lativ-a to ~h~ reaati~n mixture o~ 0,1 tC~ 5~%, preferably 3 to ~096 w~v, a~: w~l a~ with copolymeriza~le additives o~ ~ concentra~ion relativ~ to th~
~ea~tion ~olution of 0 ~o Z0% t prePer~bly 1 t~ 5~ wJ~,with crosQlinker~) o~ a concentration relatlve to the r~action mixtur~ o~ 0-5~, pre~orably 0.1 to 1% w~v, then -- optionally a~ter puri~ication -- the micropartiale~ thu~ obtaine~ are reac~d with an ~qu~ous solutio~, which contain~ --r~la~iv~ ~o ~he ald~hyd~ group co~ent o~ the mi~ropar~lGl~3 -- up ~ul~olar amount~ o~ c~upllng agen~ a~ well ~s 0 ~o ~0%, preferably 0.01 to 10~ w/v ~ur~actant~) relative to th~ total volu~., wi~h ~ rin~ up t~ 3 days, prefe~ly up to 2 d~y~, at temperatur~6 ~$ 0¢ to ~0¢, preferably 5C to 30~, at a pH o~ 3 to 9, ~refera~ly 5 to ~, and -- optionally a~ter purifi~ation --these optiQn~lly are bound to biomole~ul~e or m~cromolecul~Q.
~ h~ poly~er aldehyde particle~, o~t~ined a~ter the ~irQ~
r~otio~ step, hav~ ~ldeh~de group~ on the ~ur~aa~. With thege ~ld~hyde group~ the re~c~ivn~ typl~al for aldehydes ean ~

lg ~ p~

per~orm~d (R.C. S~hulz, Kolloidzeit3chrift und Zeitschrift ~uer Polymere, lB2 (~-2~, g~ 61) Lehrbu~h der org~ni~hen ~hemi~
~Taxtb~k o~ Organio ~hemi~ry~ IlOrganl~m,~' VEB V~rlag der Wis~en~chaf~enr ~erlin, 1~4). Thu~, it i~ pos~ible to ~ouple molecule~ o~ the particle ~urfaae whlch ahange the ~urface propertie~
Example~ ~or possi~le reac~ion~ of the ~ldehyde group :
-- reductîon o alaohol - oxldatlon to acids -~ oxima~ion : -- i~ine ~ormatlon, optionally followed ~y hydrogenation and op~lon~lly then N~alXylation -- h~drazone formation, op~ionally follow~d by hydrogenation -- m~r~aptalization -- ace.t~la~ion -- di propor~ionation by NaOH (Canniz~aro rea~tlon) -;~ aldol condensationD
The ~oupling o~ moleculefi c~n~ in~ group~ on the p~r~i~los formed in th~fir~t re~ctio~ ~tep tak~ pla~o by r~action with kh~ aldehyd~ group~. In this ca~e, for exampler ~he ~4~1~wing ~x~mpl~ry ~xp~rime~t~l condi~i4n~ n~n ~ u~ed;
1000 ~ o~ poly~rol~in p~r~icle~ are ~usp~nded in ~o ml o~
: di~killed water. sooo mg D~ ~he ~ub~tanae to be rea~te~ i~ add~d to ~hi3 p~rti~le SUBpen~iQn a~d stirred ~t room tempera~ur~.
St.irrLng i~ accomplished in kQeping with ~h~ reaçtion rat~ o~

~o t~e r~a~tion; at sl~w reaction rates up to 4a hour~. The p~r~1cl~ ~u~pen~ion i~ then dialyzed (cut off lOoOO d), I~ the ~u~st.ltuent~ int~oduced, ~.g~, by the above-lndlcatad reaction~, conta1n ~p~ionally intermediately pro~scted) fun~tional group~, the l~t~e~ can be converted aaaording to proce~e~ known to one ~killed ln ~he ar~ into ~otiv~ groups ~itable ~or coupling ~o bi~molecu~e~ ~r macromol~ul~s.
Preferred ~ch groups ar~, for example, the maleimidobenzoyl, 3-~ulfomal~imld~b~nzoyl, 4~(maleimidome~hyl)-cyclohexylcarbonyl, 4-t3-~ulfo-~malelmid~-methyl)-ayclohexyl-carhonyl, 4-(p-m~leimidophenyl)-butyryl, ~-~2-pyridyl-di~hio)propionyl~, metha~yloyl-~pentamethylene~ ~mido, brom~acetyl, io~oa~etyl, 3-i~do~r~pyl t 2-hro~oethyl., 3-mer¢aptopropyl, 2-mercaptoethyl, phenylen~i~othiocya~ate, 3-aminop~opyl, benzyl est~r, ethyl ~=ter, t-hutyl es~er, amino, C1-~ alkylamino, aminocarbonyl, hyd~azino, hydrazinocarhonyl, maleimido, metha~rylamido, metha~ryloylh~r~zinocar~onyl, ~ imidamid~¢arbonyl, halo, ~ercapto, hydrazino~rim~thylenehydrazinocarbonyl, ~minodi~byl~neamido/ ~arb~nyl, ~ro~ucarbonyl, ph~nylen~diazonium, i~o~hiocya~a~e, ~emi.~arb~zi~e, th~o~mlqarhazide, l~ocyanate groupR.
A~ ~mino group ~an, for example, be ~onv~r~d into ~n i~othio yana~e ~roup ~ç~ording t~ ~e~h~ known in thA literat~re ~e.~., wi~h thiophosgene in a two-ph~t~ system, S. Sch~ma, Synthesi~ , 803~ D.~, ~ohnson, ~. Med. Chem. L~ ol. 3~, ~36~.

, 21 ~ PI

By th~ reac~ion of ~n amino ~unct `.on an ~lpha-halo~c~ m~ de ~roup ~an be gen~r~ted with a haloacetic ~cid hallde (JA~g 196g, Vol. ~0, 450a; Chem. Phaxm~ Bull. 29 (1~, 128, lg81), which, ju~;t a~, a.g., the i~hioc:y~na~q ~roup, 1~ suitahle ~or coupling to ~iomolec~le~ or ma~romolec:ule~
A~ ~ub~ituQnt, whi~h ~n ~e ~onverted in~o a ~unctional group ~uitable ~r binding ~o a. maçrOmolOc~UlQ or l~iom~leaul~, ui~bl~ amon~ o~hers are hydroxy ancl nitroben~yl, hy~roxy and GarbOXy a:lL}syl ~5 w~ 3 thioalkyl ra~i¢~ls wlth up to 20 carbon a~.om~. They are conYerted ~ccordîng to pro~e ~e~ know~ in the litQr~tur~ to on~ ~killed in thQ art ~t~hem. Phalrm. Bull. 33, 674 (1~85), Compendium of Org. ~ynthesls V~ 5, Wlley and Son~, Inc., ~louben-Weyl, Methoden der orgahi~c:hen ~hemle ~M~thods o~
Orge~nic C:h~ try], Vol . VI~ eor~ Thleme Verlag, Stuttgart, J, ~i~c:hem~ ~,1413, (1982) ~ into the de~i.red substitu~nt~ ~fox example, with amino, hydrazino, hydrazinocarbonyl, epoxide, ~n~yd~id~, meth~cryloylhdr~zinoa~rb~nyl~ maleimidamidocarbonyl, h~lo~ haloc~xbonyl, mæ.r~apto, i~othiiocyanat~ group a~ functional grou~), and in the cas~ o~ th~ nitrob~nzyl radical ~ t a cataly~c hydrogana~on (~or example, ~aording to P.~ ~yland r, ~alytic Hydrog~n~ti~n o~r Platinum Metal~, A~ademic Pr~
1~67) to th~ ~min~benzyl derlY~tive ~an be per~o~med.
~ x~ple~ f~r the aonver~ion o~ hydroxy or amino ~roup~, bound on aromatic or ~lipha~o radiaal~, ~re ~h~ reaction~
per~ormed ln ~ui~a~le ~ol~ent~ 9uah a~ t~trahy~ro~ura~, dimethoxyethane or dimethyl sulfoxide, two-pha~ aque~u~ ~y~tem6, 3~

such .lS, e.g., water/(~ orom~tllane, in t~le presence of an acid tr~p such ~s, for example, sodium hydro.Ylde, sodium hydride or alXali or alkaline-earth cur~on~tes such ns for example, so~ium, magnesium, potassium, calcium c.~ n~tt~ or t~oly~ vinylpyridlne) l~ei]le~ ) at temperature~ ~etweell 0~ n~ the boiling point of -the re~pective solvent, bn-t pr~ferably ~et~en 20C and 60C, with a substrate of gen~ral formula 1 Nf-L-Fu (I), in which Nf is a nucleofu~e such as, e.g., Cl, Br, 1, C~l3C6H~S~3 or C~'3S03, L sta~s for an aliphat1c, ~romlltic, ~rylaliphatic, branched, straight-chain or cyclic hydrocarbon radical wi-th up to 2~ carbon atoms a~d Fu stands for the desired functional group, optionally in protected form (DE-OS 34 17 413).
As examples for compounds of general formula I there can be mentioned BrtCH2)zNH2, Br~CHz)30H, srcH2coocH3, 8rCH2C02 Bu, ClcH2coHHNH2, Br(CH2)4C02C2H5, BrCH2COBr, B~cH2coNH2~ ClCH2COOC2H5, o / \
2 2' z CH CHz, CF35031C112)38r, BrCR2C CH, BrCH CH=CH
BrCH2C6H~ Ncs, ~; Conversio~s of carboxy groups can be per~ormed, for example, accoxding to the carbodiimide method (Fiessr, Reagents for Organic Syntheses 1OJ 142), by a mixed anhydride [Org. Prep.
Proc. Int. 7, 215 (1975) or by an activated ester (Adv. Org.
Chem. Part B, 472).

.

The microparticles carrying couplln~ a~ent~ -thu~ obtained can also }: e link~d -~o hiomol~cula~ or m~cromolecule~ of which lt i~ Xno~m tha~ th~y ~p~cially concentrate in the ~rgln or o~gan p~rt to be exa~lned. Suoh molecules are, for example, en2ymes, hormones, p~ly~accharides such aR dextrans or starche~, porphyrin~, bl~omycins, in~ulin, pro~taylandin~, steroid ho~mon~ mino ~ugars, am~no acid~, peptidQ~ ~u~h as polyly3ine, protein~ t~UCh ~3, ~or example, immunoglobulin~, ~onoclonal ~ibodies), lectin~, lipid~ ~al~o in th~ ~orm ~f liposcme~) and nucleo~ide~ o~ DNA and ~NA type. Especl~lly to b~ empha3iz~d ar~
aon~ugat~s wi~h albumins, ~uch a~ hum~n ~eru~ albumin, antibodi~ u~h as, ~or ~xample, monoclonal anti~o~ies or ~,nti~yosin ~pecific for ~umor-a~o~iat~d antigen~. In~tead of blologi~al macromolecules suitable ~y~t.hetic polymer~ such as polye~hylenimine~, polamides, polyurea~, polyether~ ~uch a~
polyethylen~ ~ly¢ols and polythioureas c~n be linke~ on~ ~he ph~rm~c~ut~oal ~gent~ f~med from ~h~m are ~ultable, ~or example, ~ox u~e in tumor and in~arat di~gn~ a~ w~ll a~ tumor therapy.
Mono~lonal an~ibodies (~or ~xa~pl~, Na~ur~ ~56, ~95, 1g75) in ~ompa~ison with polyalon~l an~i~od~e~ h~v~ the advan~ge~ that they ar~ specific ~or an antigenio ~etermin~nt, have a defin~d binding ~ffini~y, ar~ hom~geneou~ (t~ heir pure representation ~ecom~ ~u~t~nti~lly impl~r) and aan be p~oduced in cell culture~ in large am~un~. As ~uch, ~or example, m~noalonal ~n~ibodio~ or thelr f~ment~ Fab and F~ab~) 2 are ~uita~le for ~u~or r~pre~entatlQn, which, for exampl~, are spe~lfl ~or human 2 ~ 3 ~

-tumors ol the gastrointestinal -tract, t~le brea~t, the liver, the bladder, the ~eproductive glands and of melanomas [Cancer Treatment Repts. 68, 317, (198~1), Bio. Sci. 3~1, 150 (198~)~ or are directed against carcinoembryonic antigell (C~A), human chorionic gona~otropin (beta-llCG) or other tumor-position arl-tigens, such as glycoproteins [New I.n~l. J. ~led. 298, 138~, (1973), US-P 4,331,647]. Antimyosin, anti-insulin and antifibrin antibodies (US-P 4,036,945) ~mong others are also suitable.
Colon carcino~as can be diagnostically detected with microparticle conjugates with antibodies 17-lA (Centocor, USA).
In the case of the antibody conju~ates the binding of the antibody on the micropartlcles must not leacl to the loss or the reduction of the binding affinity and binding specificity of the antibody for the antigen. This ca~ take place either by binding on the ca~bohydrate portion in the ~c part ol the glycoprotein or in the Fab or F(ab')2 fragments or by binding on sulfur atoms of the antibody or the antibody fragments.
In the first case, first an oxidative cleavage of sugar uni:ts for generation of formyl groups able to be coupled has to be performed. This ox.idation can be performed chemically with oxldants such as, e.g., perlodic acid, sodium metaperiadate or potassium metaperiodate according to methods known in the literature (for example, J. Histochem and Cytochem. 22, 1084, 1974) in aqueous solution in concentrations o~ 1 to 1003 preferably 1 to 20 mg/ml, and a concentration of the oxidant between 0.001 to 10 mmol, preferably 1 to 10 mmol, in a pH rangs ~' .

~s of about 4 to 8 a-t a temperature ~etween 0 an~ 3~~ ~Ind a reaction period betweel~ 15 minutes an~ ~l hours. The o~ida~ion can also be perfolmed en~ymatic.~lly, for e~am~le uit~l ~alactoseo~idase, in an enzyme conc~n-tr~ltion o~ lO-100 nnit~/ml, a substrate conr,entratioll of 1 to 20 mg/ml, ~t a pll of 5 to 8, ~
reaction period of 1 to ~ hours and a tempelature b~t~e~n 20 and ~10C (for example, J. Biol. Chem. 234, 445, 1959).
~ licropar-ticles with suitable functioncll groul,~, such as, ~or example, hydra~ine, hydrazide, hydroxylarnine, phenylhydrazine, semicarbazide and thiosemicarba~ide, by reaction between 0 and 37C, in a reaction period of l to 65 hours, a pH between about 5.5 and 8, an antibody concentration of 0.5 to 20 mg/ml and a molar ratio o~ complexing agent tô antibody aldehydes of 1:1 to 1000:1 are bound on the aldehydes generated by oxidation. The subsequent stabilization o~ the conjugate takes place by reduction of the double bond, e.g., with sodium borohydride or sodium cyaDoborohydride; in this case the reducing agent is used in 10- to 100-fold excess (for example, J. Biol. Chem. 254,4359, 1979)-The second possibility o~ formation of antibody conjugatesstarts from a gentle reduction of the disulfide bridges of the immunoglobulin molecule: in this case, the more sensitive~
disulfide bridges between the H chains of the antibody molecule are cleaved, while the S-S bonds o~ the antigen-binding region remain intact, so that practically no reduction of the binding affinity and specifici-ty of the antibody occurs ~Biochem. 18, ~6 -` 2~3~ ~7 ~226, 197~, IIalldbook of Experimental Immunology, Yo]. 1, Secolld ~dition, Blackwell Scientific Publications, I,ondon 1973, Chapter lO). These free sulfhydryl groups of ~he inter-H-chain regions .Ir. then reacted with suitable func-tional groups of the n~icroparticles at O to 37C, a p~l of a~out 4 to 7, and a reaction ~ riod of 3 to 72 I]ours with rormation of a covalent bond, which lo~s not affect the antigen binding region of the antibody. As suitable reactive groups there can be mentioned, for examp]e:
haloalkyl, haloacetyl, p-mercuric benzoate, isothiocyanate, thiol, epoxide Kroups as well as groups, which are to be ~subjected to a Michael condensation reaction, such as, for ex~mple, maleinimides, methacrylo groups (for example, J. ~ner.
Chem. Soc. 101,3097, 1979).
For linking of the antibody fragments with the microparticles there is ln addition a series of suita~le bifunctional "linkers" also commercially available (see, for example, Pierce, Handbook and General Catalogue 1~86), which are reac*ive both -toward the SH groups of the fragments and toward the amino or hydrazino groups of the microparticles.
As examp]es there~can be mentioned:
m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) ~ m-maleimidobenzoyl-N-sulfosuccinimide ester (sulfo MBS) ; N-succinimldyl-[4-(lodoacetyl)~-amino]benzoic acid ester (SIAB) ~ Succinimidyl-4(N-maleimidomethyl)-cyclohexane-l-carboxylic acid ~SMCC) ~7 a~ihimidyl-4 ~p-m~l~imidoph~nyl)-butyric acid e.3t~r (SMPB) N-~uacinlmidyl-3- ( ~-pyridyldithio) -propionia acid e~ter (S~P~) 4-[3-(2,5-dioxo-3-pyrrolinyl)-propionyloxy]-3-oxo-2,5-diphenyl-2,3-dihydrothi~phene-1,1-dioxide ~ tyl~lanyll~u~ylalanylamin~-p-b~nzyl ac~3tamido-p-tllioureido~enzyl.
A1~Q l~ond~ ~f nvn~oval~n~ typ~ ~n ~ us~d for cc~upling c~n thR hiomol~cule or macromoleaule, ~nd l~oth ionic and van der Wa~ nd hydrog~n bridg~ bond~3 in alt~rnating portion~ ar~d s~rength ~ lock and key principle~ can c:on~ribu~e to the l~inding ~for ax~mple~ avidin-l~iotin, an~i})ody~antigen). A1~ in¢lusion ao~np~unds (ho~ t) Q:E ~3mall~r complex~ in larger cAvities in ~he~ JnA~romolscule are po~ibl~.
~ rhe coupltng pr~naiple a~n~ t~ ~lr~t ln producirlg a bifunctional maàromolec~ e~ by either an antibo~y hybr.idoma, direot~d ~gainst ~ ~umor ~ntigen b~ing ~u~d wi~ a ~e~nd an~ibody hybridoma dirçot~d ag~n~ ~he miçropar~iol~ cording to t~e inventlon ~ the two ~n~ibodie~ ~elng chemlaally linked wi~h one ~noth~r hy a lin~er (~or examp~, in ~he way i~diaated i~ J. A~er. Ch~m. Soç. ~, 3097 (197~)) or the antibody, dlr~ct~d a~in~ th~ ~umor antlg~n, optionally bindlng on avl~in (or biotin) by a link~r [D.~. Hn~towi~h ~t al., J. Nu~ d. 28, 1294 (ls~7~J. Instea~ of th~ antibo~ , their Gorr~ponding F(ab~ or F(~')2 f~ nen~ oan ~e u~ed. For the pharmaoeutioal u~e, fir~ ~he b.tfunational maaromol~aule i~ lni~t~, which 2~ ~a~3 c~ncentrat~s on the t~rget ~ite, and then periodically the ml~rop~rticle ~ording t~ the invention ~optionally bound on biotin ~or avidin~], which in vivo are ooupled on the targe~ site ~nd their diagno~ic or therapautia a~tion can devel~p the~e.
Further, al~o other ~oupling method~ ~an ba used ~uch as, for ~xample, the "Re~ersible ~adiolabeling" de~ribed in Protein Tail~rlng Fo~ Me~. U~e~ [Am. ~hem. Soc, Symp.] ~1985), 34g.
With the ~o~oalled s~lid ph~s~ c~upling a par~icularly simple method for the production of antibody aon~ug~t~6 or antibody ~ragment conjugate~ vaila~ the antibody is ~ou~led on a ~ta~ionary pha~e (e.g., an ion ex~h~g~r), which i~
loaa~ed, ~or exampl~, in a glas~ aolu~n. By suaae~ive rinsing o~ th~ column wi~h a ~olution sul~able ~or genera~ion o~ ald~hyde group~, w~hing, rin~ing with a ~olu~ion of the function~lized micxoparticle~, wa hing and ~hen ~luting of ~ne ~onjugate very hlgh ~onjugate yield~ are ob~ained.
Thi~ pro~efis allow~ the a~t~matic and con~inu~u~ produc~ion o~ ~ny ~mounts o~ conjugate~.
other c~upling ~teps a~n also bR per~orm~d ln ~hi~3 way.
Thus ~ragment ¢onjugate~ aan be produc~d, for ex~mple, by the ~e~Uenc~ papain reduction/bi~no~ional linker/func~ionalized m~cropar~cl~.
~ hen the co~pound~ ~hU~ ~ormed ~r~ pr~Rr~bly chromatographic~lly puri~ied.
Particl~ in the ~ o~ 0.040100 ~i~ron~, pr~ra~ly 0.1-~ micrbnsJ ~an be produced~ ~he siz~ of the par~ Gan be ba~ally influenc~d, perh~ps wi~h a ~ew routine ori~nta~ion experiment~3, by variatlon o~ the ~t~rting concentrat~ on ol~
mono~ner, ~ux~ac~a~ n~ pH.
J3xample~ for th~ produc~ian o~ par-kicl~ f ~peci~ic ~iz¢:
1) acrol~in concentr~k~on: 10% ~wJvJ
~uri~c~an~ ~noqntration 1 . 5~ (w/v) pH: 10 . 0 ~emp~ratur~: 4~
If ~he~ conditions a~e ~alected, particle~ ars obtained wi~h an ave~rag~e diame~er ~f 750 nm.
a~ aarolein ~onc:en~ration: 20% (w/v) ~ur~aotan~ ~onoentra~lon ~ 2 (w~v) P}~' 10. 0 temperature: 2C
Under the~e ~:onditions, particl~s ar~ ob~ned Wit~ an average diams~er of 40 micrbn~
3 ) Under th~3 3am~ conditlon~ a~: under 2 ) but at a acrolein 40nl:~ntratlon o~ 109~ ~w/v~, par~icle~ ar~ o~ain~ad wi~h an ~v~rage dl~meter ~ ~ mlaron~:.
4) acroll3in conc:0ntration: 10% ~w/~
su~aCtant aonc:entration 0 . 5 ~w/v~
pH: 11.0 - ~ avera~e particl~ diame~r: 60 n~n 5) Und~r ~he aame conditions ~ und r 4) but at a pH o~ g, n aver~ge particle ~ize o~ 3 ~ 2 microne~ ~e~ul~e.
Polyglu~ral~ehyd~ ~odium hydrogensul~ite ~d~uat ~PGL) i~
us6ad as su~f~t~n~.

- 2 ~

PolyacroleiIl sodill~n hydrogeIlsulfi-te ad(luct (I~C-S03) can also be used instead of polyglutaraldehyde sodium hydrogensulfite adduct (PGL) without an effect on the particle size being observed.
Synthesis of PGL:
A 25% aqueous solution of glutaraldehyde is purified on activated carbon. Then the solution is freed of 2 by passing N2 in the aqueous solu-tion. Further, a burfer ~ollltioIl (pllosph~lte buffer, 1 molar) is adjusted to pH = 11. 1'he ~uffer solution is also freed of 2 by passage of N2. The bu~er solution and glutaraldehyde solution are brought together and polymerized under N2 atmosphere for 72 hours. Then the polymer is filtered and washed with acetone and water. The washed polymer is dried in the vacuum-drying oven at 45C. 5 g of polyglut~raldehyde is dissolved in 30 ml of H20, which contains 12.5 g of NaHS03. The solution is dial~zed with distilled H20. Then the solu-tion is freeze-dried.
The particles according to the invention can be suspended in aqueous solutions without aggregation o~ the particles occurring.
For the production of a galenic formulation, which can be administered parenterally, aqueous solutions can be used, which contain isotonizing additives such as sodium chloride, sugar alcohols (mannitol, sorbitol, xylitol, etc.) or sugars (glucose, ~ructose~. For adJustment of the p~I, buffers such as trometamol/HCl, cltric acid/NaOH, etc. can be selected.
Synthesis of PAC-S03 2 ~ 3 ~

~ . 100 ml (>f ~istille~ W~tel' i~ e~ to a three-neck flask provided with dropping funnel and stirrer an~ freed of nitrogen by passage of oxyge~. Tllen 1.829 g of K2S,,O8 is added ~nd dissolved. After the l~2S2O8 has completely dissolved, freshly distillecl aerolein is adcle(l to this solution. Then 1.14 g of AgNO3, dissolve~ 5 m] of w~ter, is added arld polymerized for 2 hours with s-tirring. The precipitated polymer is filtered off, washed several -times with wa-~er and then, to remove silver ions, 1 liter of water, in which 1.63 g of sodium thiosulfate is dissolved, is res-lsl~erJdt?cl for 1 hour. l'he polymer is filtered ~off and dried in a vacuum drying oven at 45C. The d~ied polymer lS coarsely ground in a mortar. 10 g of ground polymer is dissolved in 100 ml of sodium hydrogensulfite (37%). Then the solu-tion is dialyzed ~gainst distilled water (cut off 5000 d).
The dialyzate is used as surfactant i`or the production of polyacrolein microparticles.
B. 100 ml of dis-tilled water is placed in a round-bottom flask. 20 ml of freshly distilled acrolein is added w1th stirring. Then the pH of the reaction mixture is adjustecl to :
10.5 by addition of NaOH solution (2N) and it is polymerized for 2 hours with stirring. The preolpltated polymer is filtered off, washed several times with water and dried in a vacuum-drying oven at 4SC. 10 g of polymer is dissolved in 100 ml of NaHSO3 so~lution (37%). The solution is dialyzed against bidistilled water lcut off 5000 d). The residue is used as surfactant in the production of polyacrolein microparticles.

, .

:
..

3~
Q ~

1) In lO() ml o-f a formulation ~here ar~ contain~d p~r~iclR~: lOO mg trom~tamol; 2 ~ 4 mg + ~ICl ~or p~ = 7.4 mannitol: 5500 mg wa~r to: loo ml z~ In loo ml of a ~oxrmlla~ion there ar~ cont;ained;
par~i~le~: 50 mg ~;odlum ~hlo~ de: 360 mg wa~er ~o: loo ml A ~loating of tha pax~lcle~ ~an be prev~rlted by th~ av~rage ~en~ity of the particl~ b~inS~ adapted to that o~ the surrounding vehicl~ .
~hi~ ~an be aahieved by ad~ltion:3 o~ ~3u~tan~s of higher den~ity (x~ray contra t media, ma~nstite~. This po~ibi~ity ir-o~f~r~d ~spe~ially ln ~he ~e o~ p~t~ cle# with slight polyaldehyde s:ontent.
ThQ pharmacQutical agents aaaording to the ~ nYention contain 0~ g~lOO mg o~ micropar~icl~/ml, pre~erably ln ~g-1 mg of mi~ropar~icl~ml o~ gal~nic: ~ormulation an~ are me~ered a6 i~
rule in do~ o~ O ~ 01 ml~10 ml~X~, pre~rably O .1-1 ml/kg o~ body we~ght, e.g., $or use as ul~r~sound contra~t media. Th~y 2~re ~n~ended for enter~l and parenteral admini~tr~$on aon~ntionally; ~or example, ~or h~ar~ imaging, the imag~ ls ~ken i~nediate~y after or ~ren durin~ ~dminis~ ion c~ the contras'c ag~nt.
For u~e ln hyp~rth~rmy t:herapy the p~armaceut~c:al agents acc~rding to th~ inqre~on a.6 a rule are u~d in amount of O.~O~-îO mg, pre~r~bly 0.01-1 mg p~r g G:E ~umor.

Wlthout ~urther elaboration, it i~: ~eliev~d that one ;killed in ~hR art c!an, u~ing the pr6~cedlng de~cription, utiliz~ the present inv~n~ion to its fullest exterlt. The followlng preferred epeciflc: emb~dimen~s are, ~herefor~, to bQ ~onstrued a~ merely -~3 2 ~

illu~trative, ~n~ n~t limi~a~ive o the rem~inder ~ the disclo ure in any w~y what oever.
In th~ for~going and in ~he following ex~nlple~, all ~mp~ra~ure~ ~re ~et for~ll unoorxe~ted in d~g~ee~ Cel iu and unle~ otherwi~ indicate~, all par~s and pRrceTttage~ ~re by weight.
The entlre dl~closure of all application~, patent~ and pu~lloatlons, cited above ~nd below, ~nd of aorre~pondin~ German application P 40 04 430.0, are h~reby incorporated by referenc~.

~Ltoa~ ~or the proauotio~ o~ the oo~tr~t m~d~uat ~Qa~ti~n ~t~p:

A) An ~ueou~ solution ~ont~tining a ~uxfaatant (o~ 01-5~
w/~ aooled to OaC with atirrillg. At the ~ame time a gas is pa~d in th~ ~olution. The pH of the ~olution is adiu~tsd with Na~ to ~ha de~ired v~lu~ (pr~Rrably 9-13 ) . ~he monomes or mon~mer mlxture iB added ~o ~hi~ ~olution. The ~tirring spaed is rRduca~ a~ter 30 minutes. Aft~r ~ hour, the reac~ion ~ixtur~ is dilut~d with thQ ~bov~-in~a~ed ~u~fac~ant-containing aqu~ous ~lution. Th~ ~ixring ~p~ is redu~ed ~tlll ~o~e~ Aft~r 4 hou~s, the pre~lp~te~ mi~roparticl~, no~ ~on~ainin~ ga~, are decanted ~rom the r~maining ~uspenslo~ ~nd di~c~e~, The dec~nt~d su~ponsion i8 di~lyzs~ ts puri~y ~h~ ~ontra~ medium o~
r~iduAl m~nomers.
Yi~lds 80-~0%
~ ) An a~ueous ~o~u~ion, which c~n~ins ~hQ d~ixed surfac~ant ~nd monomer amou~, i5 ~o~Ied ~o 0C. At the same ~ime, ~ de~ire~ g~ p~s~d through tha ~nlution wi~h ~tirring. Then the pH of the ~olution iB pr~f~r~bly ~d~u~ted t4 ~ with NaOH~ ~h~ re~a~ion mix~uxe 1~ dilute~ a~t~r 1 hour.
Afte~ 3-~ hour~ th~ pen~ivn ~ont~inin~ ~h~ mioroparti~le~ i~

3~ 2~3~

separated from -the precipitated polymer whicll is discarded. The suspension is puriIied by dialysis.
Yield: 80~90%
Fxample 1 91 ml of 0.S% a(lueous surfactallt solu-tion is put into a flask. The pll o~ the solution is adjus-ted to tl with 0.2 N NaOll solution. N2 is passed through the solution. 9.5 ml of freshly dis-tilled acrolein is ins-tilled into t~e 0.5% surfactant solution cooled to 0C. Another 100 ml oI 0.57~ surfactant solution is added ko the reaction mixture af-ter 1 hour. After 3 hours the suspension containing the microparticles is decanted from the precipitated polymer and purified by dialysis.
Example 2 82 ml of 0.08% aqueous surfactant solution is put into a flask. The solution is cooled to 0C. 18 ml of freshly distilled acrolein is added to the cooled solution. Argon is passed in the solution with stirring. A-fter 1 hour, the pH of the solution is adjusted to 12 with 0.2 N NaOH solution. After 2 hours 100 ml of 0.08% surfactant solution is added. ~fter 3 hours the solution is decanted and dialyzed.
Example 3 70 ml of 0.08% aqueous surfactant solution, which contains 10% dimethylformamide, is put into a flask. The pH of the solution is adjusted -to 11.5 with 0.2 N NaOH solution. The solution is cooled to 0C. At the same time, N2 is passed in the solution. 30 ml freshly distilled acrolein is instilled into 2 ~

this solu-tion After 1 hour, ~00 ml of 0.08% surfactant solution is added -to -the solu-tion. After ~ hours the suspension is separated from the precipitated polymer and ~urifie~.
Example 4 91 mm o~` 0.5% a~ueous surfactant solution, which contains 5 rnagne-ti-te, is coo]ed to 0C in a flask. The pII of the solution is adjusted -to 12 with 0.2 N NaOH. N2 is passed through the solu-tion. 9 rnl o Ireshly distilled acrolein is instilled into the solu-tion cooled to 0C. After 1 hour, 100 ml of the 0.5%
surfactant solution is added to the reaction mixture. The suspension containing the microparticles is separated from the precipitated polymers by decan-ting and dialyzed.
Example 5 91 ml of 0.5% surfactant solution is put into a flask. The pll of the solu-tion is adjusted to 12 by addition of 0.2 N NaOII
solution. The solution is cooled to 0C. Argon is passed through the solution. 9 ml of -freshly distilled acrolein, which contains 5% butyl cyanoacrylate, is instilled into this solution.
Af-ter 1 hour another 100 ml of 0.5% surfactant solution is added.
The suspension is separated from the sediment and purified.
Example 6 91 ml of 0.08% aqueous surf~ctant solution is put in a flask. The pH o~ the solution is adjusted to 10.5 by addition of 0.2 N NaOH solution. The solution is cooled to 0C. N2 is passed through the solution. 9 ml of ~reshly distilled acrolein, which contains 20qo alpha-methylacrolein, is instilled into this 3~;
~3~

solu~i~n~ A~te~ :1 hour, ~noth~r ioo ml ~f o . 08~6 surf~ctant ~lu~ion is added. After 2 hours the miGrQ.sphere su pen~;ion i separate~ ~rom the sedim~nt and purified.
~'xample 7 ~1 ml uf 0. VR~ a~ueou~ ~ur~c~allt solutiol~, which con~in~
~5% iE;opentan~ put ln ~ îlu.~k. The $olutior~ zooled t;c ~". ~3 ml ol ~r~;hly ~lis~ Lc~ole~ added t~ t~
~;o:l~u~ion with ~3tlrrJ~ng. The ~eactlon mlxtu~e i~; filtèred ~It~r llo~lr~, Tb~ Inicrop~I~icle~ are purifled by waRhlng with wat~r.
l`he micro~;~h~r~ re re6u~3pended in wa~er.
~ urfactan~ ~ollltion: pulyglu~Lr~ yd~ ~o~llum hy drclgen~u 1 f i t~ adduct .
2nd Reaction ~
A ~u~pen~ion ~f po~ yncrolein mi~ropartiGl~ in diE~til~d t~r i,~; ~d~ t~d to a pll of ~,5 by Addltion of 0.01 N ~11 c~lu~ioll. An ~xc~ of the ~mine~con1;ainin~ lig~l~d i5 ~dded to thl~i U~lspen~ion wlth ~tirritl~ at. rot~m t~mperature. Th~ p~ o~ -l,hi~ )lutlon i~ ~re~lou~ily ad.~us1:~d to 8 by ~L~d~tlon o~ 0.01 N
N~.n~l svlution.
~ u~ u~ltly, ~tir~lng ~ per~or~n~d, ~ a func.~ion of th~
~tirrinyr ~;pe~.l, up ~ our~ ~. room tem~erature. Plalyfiis ,ink~ ifi per~o~med to r~move t~le ~oe~ mli~e~conita~lling n~i, Op~ioltally th~ lorlned S~hli~ ba~ fi ~r13 rud~ d ~o .imln~;
l~y ~ddi ~ion ~ re~ucin~ ~gentfi .
~x~mpl e B
lOOU m~ ~f poly~;rol~ln ml~ r~p~rtie~ i f roin ~x~mpl~
rc~u~;p~d~d 1~1 50 ~nl o~ w~t~r, 1000 mg ~f 3 ~mlnopropane-l-~ulfonic ~c~ dd~d to t;hiEi ~u~;pen~:ion ~n~l stlrred ~t rc~om 36a 2~3~

emper~ure for 48 hou~ The~ th~ ~ue~enElon 1~ dialyzedwater.
Th~n 1~ i~ n~l,xed wlt;h 1~0 m~ o~ Nv.BH3CN and ~3-tirred for ~4 hour~ ~t p~1 7 . ~ . Thi~ ~usp~3nsio~ h~n ~lialy~d against ~ r .
~ ;),vti.c-llally the ~mine c~n he alkylated or ~:etylated wlth chloroac0tl~ ~cid, ac~ti~ nnbydrld~ or diglycvll~ a¢ld allhydrine.

~x ~Inp 1 1000 m~ of polyuarulelll microp~rti clefi fr~m example z 1 re~uspendçd ~n ~0 m~ of wA~er. 1~0 m~ of 3-a~inoprc~p~nc phofiph~te 1~ ~d~ie(l ~0 t)lifi ~u~p~n~ien and ~ti.rred ~ room t~mpera~ure ~or ~S hour6, T~len l;he ~qpen~lon i~3 dls.lyzed lS't: w~ rl Th~n ~.i; i~ mlxed wlth 150 mg of NaBHaCN ~n(l ~3ti~red f~ 24 hour: at pH 7 . 5 . ~rhl~ ~;u~pensi~n 1~ then di~ly:~d ~gai~t w~ter .
OptiQn~ly th~ amine ~:~n be ~lkyla~d or ncetyl~ted wlth ~;hl~rv~ tic a~ , ace'cl~, 4.nhy~1rid~ or d~ycolic ncla ~nhydr~d~.

I:xarnple 10 1000 rn~ c~f poly~.rolein mic,r~?p~rticle:; from ~x~.mple 3 is Busp~nd~d ln 60 ml o~ w~ter~ 1000 m~ of 8~minoc~:tanois. a¢id i~ udde~ t~ ~hiE; ~uspan810n and ~tirred at room t~mp~rature $~r 2~ ur~;, Then th~ ~u~;p~ænslon is ~iaïy~e~l again~t wQ~el~.
Th~n it i~; mlx6J~ wlth 150 m~ o~ N~LB~ N and s~lrr~d for 24 r~ ~t pH 7~5. Thl~; su~:~pel~sion 1~ then diQly~ed ~galnst w~t~r.

37 2~3$~

t)ptlvnal.ly the ~mlne, ~n be al~yl~ted o~ ace'cyl~tr~d w~th ehloroQaetic acld T acet-l~, aQbydrlde ~T diE~lyct~lic a~ld ~r~hydrlde .

13xample 11 1000 mg o~ polyacloleln microp~r~cle~ fIOm ~x~mple 4 i~
re~uspe~ 0 ~1 v~ ~vat~r. lOOP m~ o~ ~-aminoY~leric ~cid i~
~dd~d to thi.~ susp~nslon ~,nd ~tirrod ~t room tHmp~rature ~or 36 our~. Then th~ ~;u:;p~a~ i# di~ly2i~d a~in~t water.
Th~n lt 7 ~ ~nlx~d w~ th 150 m~ o~ Na~ N ~nd ~tirred ~o~ ~4 ~our~ ~t p}~ 7, 5, Thi~ ue;pe:l~ion 1~ en dialy~ed ~g~ t wa1:er .
Opti~ lly the ~min~ can l~f~ a.l~yL~ d or acetylated wlth ehl~roacetlc a~id, a~et~.c. anhydrlde ~ diglyt:~olic aoid anhydride.

~x~mpl~
1000 mg ~ p~lyucrol~ln micr~p~r~clos ~rom exRmple ~ i~
re,~;u~pended ln ~O ml o~ w~ter. 1000 ~g o~ n~Kluco~:e~min~
hy~ro~hlorld~ dd~3d tc~ thl~ su~pen~icrl and ~tirred a~ ruom t~mp~?r~ture îor ~0 hours. I~hen the ~u~;pen~;iu~ dialyzed ~:aillst wa~ffr.
The~ mlxe :1. wlt~ 0 mg of N~ N and ~irred ~or 24 h~u3r~ ~t p~ 7 . 5, ~hl~ ~u~pen~isn 1E; then ~ ly~ed ag~;n~t wQter .
, Optiollally the amlL~3 c~n ba alky~ d or ~,~etyl~Lted with c~hloro~ceti~c aGid, a~etic ~,nhydrlde or di~ly~olic a~i~l anhyd~i~e.

~x~mple 13 100~) m~ polya~roleln microp~rti~ s ~Fom ~x~mple 8 ~#
r~uspQnded in 50 ml ~ w~ . 1000 m~ o:~ hexame~hyl~n~dlami~
1~ &Lddocl to thl~ ~u~pens~ 7.nd ~tirrqd ~ rootn t~mJ~ral;ur~ r 24 hour-~. Then the ~uE;pen~ion 1~ dl~ly~ed ag~ln~t w~er.

T~ nixed wl~h 150 mg o~ Nal~3CN and 14t;1rr~d ~r 24 hours a~ ~l 7.5. ~hl~ su~pens1~n 1E; the~ dl~lyzed ~ n6* wator.
0~1;ic~nQlly ~he ~minc can be ~lkylat~(l or acetyla~ed ~ith ~,loroac~lc hCiC3, ac~;iç ~n~ydride or digly~ollc ac~ ~nhydr~de.

Ex~pl~ 14 lOOU mg oI' polyacrolein micrvparl~lcle~ :~rom ox~mple 7 i*
refiu~pend~?d in ~0 ml o~ w~1:er. 1000 m~ of polyly~ine (hSW ~
~S;~, f300 clal~n.~ ded to 1~ uspen~ic)n and ~tlrred ~ rvom tem~erature ~or :~0 hour~;. Th~h th~ p~3n~ion i~; w~sh~d wi~
w~ t~r .
~xnmp~o 15 10~ mg of pol~ in micropartlcle~ lrom example 5 1~
rç~u~peJl~led in 2 . ~ ml ~ YV~r . 2~ m~ o~ huma~ ~erum albumln, sol~ in 2.5 ml o~ w~t~r, is adde~ to thi~; f3u~pen6ion ~n~
~;tlrl~d ~tl; rc,om temp~ature ~or ~ hours ~ Th~n th~ ~uE:pens~on t di~lyzed against dis~tilled w~ter (~U~ 0~1 loO,000 ~x~u pl~
1000 mg of poly~crolein m~.~roparti~ rom exa~pl~ 4 1 re~usp~ncled ln 50 ml of wa'c~, 1000 mg o$ ~ ~-dlethylan~inc) ~ -~hylamin~ ~8 ~dded 1;~ tl~ fipsnfiioll a~d sti~red at room ten~pert~tu~e f~r 20 hou~, Th~n ~he~ su~3pens~0t- 1~ dialyz,~d ~in~t wat~r.
E~caJnplq 17 300 mg of polyacrolein mioropar~i~les ~ror~ ~xample 1 is resu~pended in 7.5 ml o: d.istillçd water. 750 mg of ~

. 3g~
2~3~7 The preG~ing examples can be repea~ed with ~imi.lar ~ucces~
hy ~6tituting t~e generic~lly or speci~icall~ de~cribed reactant~ ~nd~or operatiny condltions o~ this inv~nt~on ~or those used in the pr~cecling examples.
From the foxegoing de~aription, one ~kllled in the ~rt aan easily a~cert~in the e~enti~l ~haractR~lsti~s of thi~ inventlon, and without dep~r~ing ~rom ~he ~pi~it ~nd ~op~ thereof, ca~ ~ake v~rio~ changes ~nd ~o~iflcation~ he in~ention to adapt ~t to va~iou~ ua~e~ arld condition~.

~ 2~ 7 In vltro tests In ln vitro experimen-ts by backscattering measurements of the echo ampli-tude of suspensions of n~icroparticles according to the ln~ntivn, se~ected as examples, t~eir very good acoustic pro~ert~e~ are subst~n~iated.
To e.~plain the in vitro tests and the figures obtained from them:
The me~suring device consists of an ul-trasonic transmitter combined with an ~ltrasonic receiver and a measuring cuvette with the specimen. An ultrasonic pulse is sent out to measure the acoustic propertles of the specimen. The pulse is scattered on the glass wall of the cuvette, passes through the specimen and then, if the specimen is not echogenic, is scattered on the rear of the cuvette. The backscattering of the ultrasonic pulse is measured by the receiver a~d indicated by a change of the amplitude (see figures).
The backscattering behavior o~ water (as example for the nonechogenic specimen) is represented in fig. 1. The backscattering amplitude of -the front wall (a-t 3 microseconds) and the back wall (at about 16 microseconds) of the cuvette can be clearly seen.
If an echogenic specimen is measured, a backscattering behavior results as reproduced in fig. 2-4. The backscattering signal of the cuvette wall is not maintained, since the ultrasonic pulse by the interaction with the echogenic specimen 2 ~

is dissipated or so challged -that no backscattering to the receiver can any longer take place.
The backscatt~ring amplitudes of aqneous particle ~uspensions o~ e~amples 8 (fi~. 2), 11 (fig. 3) and 15 (fig. 4), i n a concel~tra-tion of O . 5 mg/ml each, w~re cl~-termined.

In vivo tests To perform an echocardiographic examination on a dog (beagle) of about 10 kg, the contrast media according to the invention were used as follows: from the vial with the ready-made suspension 1 ml of the solution is taken, which contains 40 micrograms/ml of particles coupled with albumin (example 15) in 5% glucose solution. The injection of this contrast medium takes place in the vena saphena ramus caudalis by a three-way cock, open all-around, with an injection rate of at least 1 ml/s, but more favorable with a rate of 3 ml/s, followed by a postinjection of 5 ml of a physiologically common salt solution ( 0 . 9qo) . The postinjection takes place to obtain a contras-t medium bolus lasting as long as possible. Before the injection (igo 5) an "apical four-chamber view" is put on the test animal with a usual commercial sound head f~r the echocardiography on the thora~ wall (transthoracic lead) and fastened with a clamp. Before, during and ~fter the injection the sound lead are indicated on the display of the ultrasonic examination device and opti.onally documented on videotape or with a vldeoprinter. This test ~2 2~

arrangement corresponds -to the prior art and is kno~ to one skilled in the ar-t.
With the ultrasonic contrast medium reaching the ri~ht heart, t.he contrast effects can be tra~ked in color ~oppl~r, ~n 2D echo image or in M mode echo,image. The contrast medlum fir~t labels the blood of the right atrium, then tl-e right ventricl~
and finally the pulmonary artery are contrasted. In this case, a homogeneous filling occnrs, which lasts a su~ficient time for a diagnostic examination. While the cavities of t~e right heart are again filled with uncontrasted blood (decrease and disappearance of the echogenity in the heart cavities), tbe contrast medium appears after the lung passage ~transcapillary) in the pulmonary veins, then fills the left atrium, the left ventricle and the subsequent high-pressure system homogeneously.
The contrast effects in -the cavities o~ the left heart last longer than those in the right heart side. Besides the contrasting of cavities of the left heart, a contrasting of other organs, which the blood circulation reflects, takes place.
Figure 6 shows the filling of the left ventricle with contrast medium.
The use of the ultrasonlc con*rast media according to the invention is not limited ~o visualization of ~he b~ood streams in the vessel systeml a contrasting of body cavities is also possible. Caused by tlle blood circulation representation the examination of other vrgans with these contrast media can also take place with ~ood succe~s.

:

Claims (25)

1. A microparticle comprising a biodegradable polyaldehyde synthesized from an aldehyde monomer and, optionally, other monomers and/or crosslinkers capable of copolymerization, and, optionally, a surfactant, a gas and/or highly volatile liquid in free or bound form, a coupling agent, a biomolecule or macromolecule bound by a coupling agent, and/or a diagnostically or therapeutically effective component.

2. A pharmaceutical composition comprising a microparticle according to claim 1 and a pharmaceutically acceptable adjuvant.

3. A method of artificial partial hyperthermy comprising administering to a patient microparticles of claim 1 and taking an ultrasonic image of the patient.

4. A method of artificial partial hyperthermy comprising administering microparticles of claim 1 to a patient in need of such treatment and heating said particles in said patient.

5. A method of achieving a concentration of an agent in a patient comprising administering said agent as a microparticle of claim 1 which further contains a magnetic component and applying a magnetic field to localize said microparticles in said patient.

6. A microparticle according to claim 1, wherein the polyaldehyde is polymerized from aldehyde monomers which are:
I. alpha,beta-unsaturated aldehydes, II. alpha-substituted acrolein derivatives, III. and/or dialdehydes.

7. A microparticles of claim 6, wherein said aldehydes I, II and/or III are:
I. acrolein crotonaldehyde propynoicaldehyde II. alpha-methyl acrolrin alpha-chloroacrolein alpha-phenyl acrolein alpha-ethyl acrolein alpha-isopropyl acrolein alpha-n-butyl acrolein alpha-n-butyl acrolein alpha-n-propyl acrolein III. glutaraldehyde, succinaldehyde alpha- substituted acroleins betasubstituted acroleins ethyl cyanoacrylates methyl cyanoacrylates butyl cyanoacrylates hexyl cyanoacrylates methyl methacrylates vinyl alcohols acrylic acids methacrylic acids acrylla acid chlorides methacrylic acid chlorides acrylonitrile methacrylonitriles acrylamides substituted acrylamides hydroxy methyl methacrylates mesityl oxide dimethylaminoethylmethacrylates 2-vinylpyridines, and/or N vinyl-2-pyrrolidinone.

8. A microparticle according to claim 1, containing a surfactant which is ionic or nonionic.

9. A microparticle of claim 7, wherein the surfactant is polyethylene oxide; a polyoxyethylene polyoxypropylene, a carboxylic acid salt; a polyoxyethylene fatty acid ester: sodium dioctyl sulfosuccinate polyglutaraldehyde sodium hydrogensulfite adduct, polyacrolein sodium hydrogensulfite adduct and/or polyvinyl sulfonic acid.

10. A microparticle of claim 9, wherein the surfactant is Pluronic(R) F68, Pluronic(R) F 108, Pluronic (R) F 127, polyethylene glycol, Poloxamin 908, Polaxamer 707, sodium oleate or polyoxyathylene stearate.

11. A microparticle according to claim 1, containing a gas or highly volatile liquid.

12. A microparticle of claim 11, wherein the gas or highly volatile liquid is ammonia; air; an inert gas: a sulfur halide:
nitrogen; carbon oxides; oxygen; hydrogen; a hydocarbon; a halogenated hydrocarbon; an ether; a fluorinated ether:
dimethylaminoaceton; propylene oxide: N-ethylmethylamine; N-ethyldimethylamine; and/or furan.

13. A microparaticle of claim 12, wherein the gas or highly volatile liquid is helium, neon, argon, xenon, krypton, sulfur hexafluoride, methane, ethane, propane, butane, pentane, neopentane, isopentane, oyclopentane, ethylene, propylene, acetylane, 3,3-dimethyl-1-butyne, 2,3-pentadiene, 2-methyl-2-butene, 2-methyl-1,3-butadiene, 2-butyne, 2-methyl-1-butene, 3-methyl-l-butene, methylene chloride, 1,1-diahloroethylene, isopropyl chloride, dibromodifluoromethane, bromomethane, dimethyl ether, diethyl ether or a fluorinated ether.

14. A microparticle according to claim 1, containing as coupling agent I. A compound containing an amino group, II. A compound containing an acid group, III. A compound containing a hydroxy group, or IV. A polymerizable substance.

15. A microparticle of claim 1, wherein said coupling agent is hydroxylamine, butylamine, allylamine, ethanolamine, trishydroxymethylaminomethane, 3-amino-1-propanesulfonic acid, 5-aminovaleric acid, 8-aminooctanoic acid, D-glucosamine hydrochloride, aminogalactose aminosorbitol, aminomannitol, diethylaminoethylamine, an aniline, sulfonyl acid amide, choline, N-methylglucamine, piperazine, 1,6-hexanediamine, urea, hydrazine, glycine, alanine, lysine, serine, valine, leucine, a peptids, a protein, albumin, human serum albumin, polylysine, gelatin, a polyglycolamine, an aminopolyalcohol, a dextran sulfate with amino groups, N-aminopolyethylene glycol (HO-PEG-NH2), N,N'-diaminopolyethylene glycol (NH2-PEG-NH2), an antibody, an immunoglobulin, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linolic acid, linolenic acid, cyclohexane carboxylic acid, phenylacetic acid, benzoylacetic acid, chlorobenzoic acid, bromobenzoic acid, nitrobenzoic acid, ortho-phthalic acid, meta-phthalic acid, para-phthalic acid, salicylic acid, hydroxbenzoic acid, aminobenzoic acid, methoxybenzoic acid, (PEG linker asparaginic acid), PEG linker glutaminic acid, PEG
linker DTPA, PEG linker EDTA, methanol, ethanol, propanol, butanol, pentanol, hexanol, heptanol, octanol, decanol, dodecanol, tetradecanol, hexadecanol, octadecanol, isopropyl alcohol, isobutyl alcohol, isopentyl alcohol, cyclopentanol, cyclohexanol, crotyl alcohol, benzyl alcohol, phenyl alcohol, diphenyl methanol, triphenyl methanol, cinnamyl alcohol, ethylene glygol, 1,3-propanediol, glycerol, pentaerythritol, acrolein, crotonaldehyde, propynoicaldehyde, alpha-methylacrolein, alpha-chloroacrolein, alpha-phenylacrolein, alpha-ethylacrolein, alpha-isopropylacrolein, alpha-n-butylarolein, alpha-n-propylacrolein, glutaraldehyde, succinaldehyde or their derivatives or their mixture with an additive capable of copolymerization which is an alpha-substituted acrolein, beta-substituted acrolein, ethyl cyanoacrylate, methyl cryanoacrylate, butyl cyanoacrylate, hexyl cyanoacrylate, methylmethacrylate, vinyl alcohol, acrylic acid, methacrylic acid, acrylic acid chloride, acrylonitrile, methacrylonitrile, acrylamide substituted acrylamide, hydroxymethylmethacrylate, mesityl oxide, dimethylaminoethylmethacrylate 2-vinylpyridine, or N-vinyl-2-pyrrolidone.

16. A microparticle according to claim 1, containing a biomolecule or macromolecule which is organ- or tissue-specific.

17. A microparticle of claim 16, wherein said biomolecule is a monoclonal antibody.

18. A microparticle according to claim 1, containing a diagnostically or therapeutically effective component for diagnosis or therapy of tumors.

19. A microparticle of claim 18, wherein said component is doxorubicin, actinomycin, magnetite, mitomycin C or triamcinolone.

20. A process for the production of microparticles according to claim 1, comprising reacting with mixing an aqueous solution containing 0 to 40% w/v, of surfactant(s) and 0 to 10%
w/v of diagnostically or therapeutically effective components and gases or highly volatile liquids, at a temperature of -5°C to +80°C, a pH of 7 to 14, within 1 minute to 24 hours, and optionally with the passage of gas, with copolymerizable aldehyde(s) up to a concentration relative to the reaction mixture of 0.1 to 50% w/v, a wall a with copolymerizable additives of a concentration relative to the reaction solution of 0 to 20% w/v, with crosslinker(s) of a concentration relative to the reaction mixture of 0-5% w/v, then -- optionally after purification -- reacting the microparticles thus obtained with an aqueous solution, which contains --relative to the aldehyde amount -- up to equimolar amounts of coupling agent as well as 0 to 20% w/v, surfactant(s), relative to the total volume, with stirring up to 3 days, at temperatures of 0°C to 60°C, at a pH of 3 to 9, and -- optionally after purification -- binding to the resultant microparticles a biomolecule or macromolecule.

21. A process of claim 20, comprising reacting with mixing an aqueous solution containing 0.01 to 10% w/v of surfactant(s) and 0 to 10% w/v of diagnostically or therapeutically effective components and gases or highly volatile liquids, at a temperature of 0°C to 40°C, a pH of 9 to 13, within 1 hour to 10 hours, and optionally with the passage of gas, with copolymerizable aldehyde(s) to up a concentration relative to the reaction mixture of 3 to 20% w/v, as well as with copolymerizable additives of a concentration relative to the reaction solution of 1 to 5% w/v, with crosslinker(s) of a concentration relative to the reaction mixture of 0.1 to 1% w/v, then -- optionally after purification -- reacting the microparticles thus obtained with an aqueous solution, which contains -- relative to the total volume, with stirring up to 2 days, at temperatures of 5°C to 30°C, at a pH of 5 to 8, and -- optionally after purification -- binding to the resultant microparticles a biomolecule or macromolecule.

22. A process for the production of a pharmaceutical agent according to claim 2, wherein the microparticles, dissolved or suspended in water, optionally with the additives usual in galenicals, are put in a form suitable for enteral or parenteral administration.

23. Microparticles according to claim 1, having a diameter of 0.1 to 40 µm.

24. Pharmaceutical agents according to claim 2, wherein the concentration of microparticles is 1 micrograms to 100 mg per ml of galenical formulation.

25. Pharmaceutical agents according to claim 2, wherein the concentration of microparticles is 10 micrograms to 1 mg per ml of galenical formulation.